alexa Mechanosensitive Signaling Pathways Detection During TCFA Formation
ISSN: 2332-0737

Current Synthetic and Systems Biology
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3rd International Conference on Systems and Synthetic Biology
July 20-21, 2017 Munich, Germany

Rob Krams, Takayuki Homma, Leila Towhidi, Zoltan Kis, Hugo Sant’Ana Pereira, Nataly Maimari, Nisrine Zogaib, Delaram Khodadadi, Mohammed Ahmed, Anouk Post, Emir Karadza, Michael Dent, Vikram Mehta, Anusha Seneviratne, Ranil de Silva and Ryan M Pedrigi
Imperial College London, UK
Posters & Accepted Abstracts: Curr Synthetic Sys Biol
DOI: 10.4172/2332-0737-C1-009
Abstract
A surge of recent studies have highlighted the role of blood flow in determining plaque growth and plaque composition, both in animal studies and recent clinical trials. Despite these compelling data, the underlying mechanism for flow-induced plaque formation is currently unknown. Blood flow is sensed by endothelial cells, and these cells react to blood flow by changing their gene signature. Initial studies showed that approximately 2000 genes are regulated by blood flow, distributed over 40 signaling pathways regulating 15 transcription factors and their changes during plaque development are currently unknown. In this presentation, we report the development of a novel platform that enables to study the adaptation of endothelial gene networks in vascular biology. This platform consists of state-of-the-art ultra-high resolution, small animal imaging (μCT, μMRI and US) coupled to finite element methods to determine the endothelial shear stress and strain fields during the development of the vulnerable plaque. These maps subsequently gave a robot-driven laser-capture machine (Zeiss, PALM MicroBeam) which isolated groups of endothelial cells exposed to a certain shear and strain field during plaque development. Next, RNA was isolated, amplified with linear amplification kits, and libraries are prepared for deep RNA sequencing (Illumina HiSeq 2500). After this, in-house developed bioinformatics tools were applied to decipher gene networks and gene modules. Entire gene networks were subsequently tested with a newly in-house developed siRNA microfluidics system for cardiovascular studies, which consists of a programmable robot-dispenser and a bespoke, in-house developed combined flow-cell and microporator. In parallel to this, live cell imaging tools are developed to determine network dynamics from single cell measurements, which was coupled to the siRNA microfluidics system to steer synthetic network design. In a first series of studies, we have developed a synthetic gene network to determine the activity of a GPCR-dependent mechanosensor and we have coupled this novel technology to a microfluidic chamber to test a library of small drugs to decipher a novel treatment of flow-induced TCFA.
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Email: [email protected]

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